Ultra-performance Liquid Chromatography-triple Quadrupole Research Articles

Highly sensitive determination of three kinds of amanitins in urine and plasma by ultra performance liquid chromatography-triple quadrupole mass spectrometry coupled with immunoaffinity column clean-up

建立了超高效液相色谱-三重四极杆质谱高灵敏测定尿液和血浆中α-鹅膏毒肽、β-鹅膏毒肽和γ-鹅膏毒肽的方法。经过免疫亲和柱净化,尿液样品浓缩20倍、血浆样品浓缩10倍,以Kinetex Biphenyl色谱柱(100 mm×2.1 mm, 1.7 μm)作为分析柱,甲醇-0.005%(v/v)甲酸水溶液作为流动相进行梯度洗脱分离,电喷雾电离、负离子、多反应监测模式下检测,外标法定量。3种鹅膏毒肽的线性范围为0.1~200 ng/mL,相关系数(r)>0.999。尿液和血浆中3种鹅膏毒肽的基质效应和提取回收率分别为92%~108%和90%~103%,变异系数均小于13%。尿液中3种鹅膏毒肽的准确度为-9.4%~8.0%,重复性和中间精度分别为3.0%~14%和3.5%~18%,当取样量为2.00 mL时,方法的检出限均为0.002 ng/mL;血浆中3种鹅膏毒肽的准确度为-13%~8.0%,重复性和中间精度分别为3.9%~9.7%和5.5%~12%,当取样量为1.00 mL时,方法的检出限均为0.004 ng/mL。该法操作简单、灵敏、准确,已在中毒患者摄入野生蘑菇后138 h的尿液中检出0.0067 ng/mL α-鹅膏毒肽和0.0059 ng/mL β-鹅膏毒肽。该法已成功解决中毒患者尿液和血浆中超痕量鹅膏毒肽的检测难题,对于疑似中毒病人的早诊断、早治疗、降低死亡率都具有非常重要意义,也为今后开展此类毒素毒理作用及机体代谢规律的研究提供了可靠的技术支撑。

Distinct Urinary Metabolic Biomarkers of Human Colorectal Cancer.

Colorectal cancer (CRC) is one of the most commonly diagnosed cancers with high mortality rate due to its poor diagnosis in the early stage. Here, we report a urinary metabolomic study on a cohort of CRC patients (n =67) and healthy controls (n =21) using ultraperformance liquid chromatography triple quadrupole mass spectrometry. Pathway analysis showed that a series of pathways that belong to amino acid metabolism, carbohydrate metabolism, and lipid metabolism were dysregulated, for instance the glycine, serine and threonine metabolism, alanine, aspartate and glutamate metabolism, glyoxylate and dicarboxylate metabolism, glycolysis, and TCA cycle. A total of 48 differential metabolites were identified in CRC compared to controls. A panel of 12 biomarkers composed of chenodeoxycholic acid, vanillic acid, adenosine monophosphate, glycolic acid, histidine, azelaic acid, hydroxypropionic acid, glycine, 3,4-dihydroxymandelic acid, 4-hydroxybenzoic acid, oxoglutaric acid, and homocitrulline were identified by Random Forest (RF), Support Vector Machine (SVM), and Boruta analysis classification model and validated by Gradient Boosting (GB), Logistic Regression (LR), and Random Forest diagnostic model, which were able to discriminate CRC subjects from healthy controls. These urinary metabolic biomarkers provided a novel and promising molecular approach for the early diagnosis of CRC.

Serum Amino Acid Profiles Predict the Development of Hepatocellular Carcinoma in Patients with Chronic HBV Infection.

Background: The study aimed to find out the alterations in serum amino acid (AA) profiles and to detect their relationship with carcinoma formation. Methods: Targeted metabolomics based on ultraperformance liquid chromatography triple quadrupole mass spectrometry to quantitatively analyze serum AA levels in 136 hepatitis B (CHB) patients and 93 hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) patients. Results: It was shown that decreased serum levels of leucine, lysine, threonine, tryptophan, valine, serotonin, and taurine were observed in more HCC patients than CHB patients, but the serum phenylalanine level was increased. Serum valine and serotonin were lower in Class C than Class A and Class B in HCC patients. Accompanied with the higher score of Model for End-Stage Liver Disease, serum phenylalanine was increased not only in CHB patients but also in HCC patients. The serum level of phenylalanine increased in the decompensated stage more than in the compensated stage, while serum leucine and serotonin significantly decreased. Serum serotonin still had significant differences between CHB and HCC both in the HBV desoxyribonucleic acid (HBV-DNA) negative group and in the HBV-DNA positive group. Furthermore, it was shown that the tryptophan ratio, branched-chain amino acids (BCAA)/aromatic amino acids ratio, BCAAs/tyrosine ratio, Fischer’s ratio, and serotonin-to-tryptophan ratio significantly decreased, while the tyrosine ratio and the kynurenine-to-tryptophan ratio increased in HCC patients more than those in CHB. Conclusions: A distinct metabolite signature of some specific serum amino acids was found between CHB and HCC patients, which may help predict the development of HCC at an early stage.

Profile of Twenty-Three Human Milk Oligosaccharides in Han Chinese Mothers throughout Postpartum 1 year

Human milk oligosaccharides (HMOs) are multifunctional carbohydrates in breast milk, which are composed by a variety of structures. This study aimed to identified HMOs concentration profile, milk microbiota composition, and the associations with major maternal characteristics in Han Chinese mothers in the one-year lactation period. Seventeen healthy mothers aged from 28 to 36 years, who gave birth to healthy term infants, were recruited. Carbohydrates were detected using the MIRIS human milk analyzer (HMA), and twenty-three HMOs were quantified using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS). Results showed that carbohydrates were relatively stable, while total HMO concentrations ranged from 1.74 to 9.72 g/L and decreased gradually over lactation in breast milk. Based on the structure, seven sialylated HMOs concentration showed the significant decline (p<0.05) after three months in lactation. In addition, the relationships between maternal factors, containing the lactation period, genetic status, delivery mode, parity, and milk microbiota profile, and the HMO composition in healthy women, which still need further investigations, were observed.

11 Determination of Milk Concentrations and Pharmacokinetics of Salicylic Acid Following Acetylsalicylic Acid (Aspirin) Administration in Postpartum Dairy Cows

Abstract Administration of non-steroidal anti-inflammatory drugs, such as the widely available drug aspirin, or acetylsalicylic acid (ASA), may improve health and milk production in postpartum dairy cows. However, plasma pharmacokinetic (PK) data for salicylic acid (SA), the active metabolite of ASA, are unrepresentative of lactating dairy cows and current treatment regimens, and there are currently no published milk residue and minimal pharmacodynamic data available for ASA or SA in dairy cows. The objectives of this descriptive study were to (1) describe the pharmacokinetics of SA in the milk and plasma of postpartum dairy cows following oral ASA administration; (2) to estimate a milk withdrawal period for dairy cows treated with ASA; and (3) to determine the impact of ASA on plasma prostaglandin E2 metabolite (PGEM) concentrations. Primiparous (n = 3) and multiparous (n = 7) postpartum Holstein dairy cows received 2 oral treatments with ASA at 200 mg/kg, 24 h apart. Plasma and milk SA concentrations from 0 h through 120 h after ASA administration were analyzed using ultra-performance liquid chromatography triple quadrupole mass spectrometry. Plasma PK analysis was performed using a non-compartmental approach using the free software, PKSolver, in Excel. An estimated milk withdrawal interval of 168 h was determined using the FDA Milk Discard App in R (RStudio, Boston, MA) using a 99th percentile upper tolerance limit with 95% confidence. Plasma PGEM concentrations from 0 h to 24 h after ASA administration were determined in duplicate using ELISA and were reduced compared with baseline for up to 12 h after ASA administration. A mean reduction in PGEM of -49.3% was observed at 2 h. Results from this study suggest that the current milk withhold recommendation of 24 h for dairy cows administered ASA should be revised and that ASA administration may mitigate postpartum inflammation through reduction in prostaglandin production.

Quality evaluation of Tetrastigma hemsleyanum different parts based on quantitative analysis of 42 bioactive constituents combined with multivariate statistical analysis.

The root of Tetrastigma hemsleyanum (RTH) has been widely used as a folk medicine in China. Meanwhile, its stems (STH) and leaves (LTH) are consumed as functional tea and food supplementation. Therefore, it is important to get a better understanding of the distribution of bioactive constituents in different parts of T. hemsleyanum. To develop a method for quantitative analysis of multiple bioactive constituents and comparing their distribution in RTH, STH and LTH. Ultra-performance liquid chromatography triple quadrupole ion trap tandem mass spectrometry (UPLC-QTRAP-MS/MS) was used for the quantitative analysis. The quantitative data were further analysed by principal component analysis (PCA), hierarchical cluster analysis (HCA) and partial least squares determinant analysis (PLS-DA). Forty-two constituents in RTH, STH and LTH, including 14 flavonoids, three phenolic acids, 15 amino acids and 10 nucleosides, were quantitatively determined. The contents of flavonoids and phenolic acids in LTH were significantly higher than those in RTH and STH. While the contents of amino acids and nucleosides in LTH were less than those in RTH and STH. Multivariate statistical analysis can significantly classify and distinguish RTH, STH, and LTH. The present method would be helpful for the quality control of T. hemsleyanum, and the results would be useful for the efficient utilisation of T. hemsleyanum in the future.

Glycerophospholipid metabolism is involved in rheumatoid arthritis pathogenesis by regulating the IL-6/JAK signaling pathway

To explore the metabolic mechanism of differential plasma interleukin (IL)-6 expression in patients with rheumatoid arthritis (RA). A total of 240 RA patients were enrolled in the non-target metabolomics study cohort and 69 healthy volunteers were included as healthy controls (HCs). Plasma IL-6 levels were detected by electrochemiluminescence assay. Plasma metabolites were detected by ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Patients with active RA (n = 20) and remissive RA (n = 20) and 20 HCs were enrolled in the targeted validation cohort. Metabolites identified by non-target metabolomics were quantitatively analyzed by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry. Effects of 1-oleoyl-sn-glycero-3-phosphocholine (OGPC) associated with IL-6 on MH7A cells were assessed. After 24-h or 48-h induction by TNF-α, the supernatants were collected for IL-6 quantification by enzyme-linked immunosorbent assay. Furthermore, Western blot was performed to investigate the relative JAK2 and p-JAK2 expressions. With an increasing IL-6 level, OGPC shown to be related to the glycerophospholipid metabolism pathway by Kyoto Encyclopedia of Genes and Genomes analysis displayed a significant decrease. In the validating RA cohort, the OGPC concentrations in remissive RA group and active RA group decreased compared with HC group. OGPC down-regulated IL-6 secretion and p-JAK2 expression in TNF-α-induced MH7A cells in vitro. In conclusion, glycerophospholipid metabolism is the main metabolic pathway associated with the differential IL-6 expression in RA patients. The down-regulated OGPC is a promoting factor for the increased IL-6 plasma level in RA patients, which further affects the downstream JAK signaling pathway.

Simultaneous determination of 36 hypotensive drugs in fingerprints by ultra performance liquid chromatography-triple quadrupole composite linear ion trap mass spectrometry

指印中蕴含着供体摄入成分等相关信息,通过对其分析可对供体进行特征刻画,从而为案件侦查提供线索,指印分析也可用于药物摄入的定性检测,因此检验指印中的降压药具有重要的实际应用价值。建立了超高效液相色谱-三重四极杆复合线性离子阱质谱(UPLC-Q-TRAP/MS)同时检测指印中36种降压药的方法。前处理方法采用沉淀蛋白法,使用3×3 cm滤纸采集指印,将滤纸剪碎置于2 mL塑料离心管中,加入0.50 mL甲醇,涡旋混合1 min,超声振荡3 min,取出后以12000 r/min离心5 min,取上清液进样分析。采用ACQUITY UPLC® BEH C18柱(100 mm×3.0 mm, 1.7 μm)分离,以0.01%甲酸水溶液和甲醇作为流动相进行梯度洗脱。质谱分析采用可编程多反应监测-信息关联采集-增强子离子(SMRM-IDA-EPI)扫描方式,在高灵敏度分析的同时进行二级谱库检索,增加定性结果的准确性。36种药物在0.05~50.00 ng/fingerprint范围内线性关系良好,相关系数(r)大于0.99,检出限和定量限分别为0.001~0.045 ng/fingerprint和0.002~0.050 ng/fingerprint,在0.25、2.50、25.00 ng/fingerprint 3个加标水平下的基质效应为79.0%~119.2%,回收率为79.3%~116.2%,日内精密度为0.2%~18.3%,日间精密度为1.6%~19.1%。使用该方法检测了87名高血压患者指印中的降压药,大部分样本中可检出其所服用的药物。该方法操作简单,灵敏度高,选择性好,适用于指印中降压药的筛查检验。

Fast screening of tyrosinase inhibitors from traditional Chinese medicinal plants by ligand fishing in combination with in situ fluorescent assay.

A simple and rapid method for screening of tyrosinase (TYR) inhibitors present in traditional Chinese medicines (TCMs) was developed by combining ligand fishing and the fluorescent enzymatic assay based on dopamine-functionalized carbon quantum dots (CQDs-Dopa). Ligands of the enzyme present in the TCM extractions were firstly adsorbed on the enzyme-modified magnetic beads, and then the beads were magnetically separated and subjected directly to the CQDs-Dopa-based fluorescent assay. Finally, compounds were desorbed from the "active" beads and identified with ultra-performance liquid chromatography-triple quadrupole mass spectrometry. A known natural TYR inhibitor quercetin was selected to assess the feasibility and quantification performance of this method, and good linearity in the range of 0.01-0.16mM (R2 = 0.992) with a low detection limit of 0.004mM was obtained. This method was then applied to screen TYR inhibitors present in Scutellaria baicalensis and Sophora flavescens. Six TYR inhibitors including baicalin (1), baicalein (2), wogonin (3), oroxylin A (4), kurarinone (5), and sophoraflavanone G (6) were found, among which 1-4 were firstly discovered in this work. This is the first report on the in situ assessment of the target compounds obtained by ligand fishing in the form of a mixture, which exhibited the combined advantages of specific extraction ability of ligand fishing and the high sensitivity of CQDs-based fluorescent assay, showing great potential for fast screening of enzyme inhibitors from TCMs.

Targeted Metabolomics Reveals Birth Screening Biomarkers for Biliary Atresia in Dried Blood Spots

Early diagnosis and timely surgical Kasai portoenterostomy greatly improve the survival of patients with biliary atresia (BA), a neonatal cholestatic disease, which has encouraged investigators to develop newborn screening for BA. In this study, we used ultraperformance liquid chromatography-triple quadrupole mass spectrometry-based targeted metabolomics profiling to identify potential BA biomarkers in dried blood spots (DBS) collected from BA patients (n = 21) and healthy controls (n = 100). A distinctive metabolic profile comprising eight significantly differentially expressed metabolites, taurohyocholic acid (THCA), glutamic acid, 2-hydroxyglutaric acid, ketoleucine, indoleacetic acid, alpha-ketoisovaleric acid, glycocholic acid, and taurocholic acid (TCA), clearly distinguished BA infants from control neonates. Three metabolites, THCA, 2-hydroxyglutaric acid, and indoleacetic acid, were selected using linear regression and receiver operating characteristic (ROC) curve analysis and model construction. The area under the ROC curve for this model to discriminate between BA and comparison infants was 0.938 (95% confidence interval, CI: 0.874-1.000). A cutoff value of -0.336 produced a sensitivity of 90.48% (95% CI: 69.62% - 98.83%) and specificity of 92% (95% CI: 84.84% - 96.48%). In conclusion, the results suggest that metabolic markers in DBS obtained from newborns have a great potential for BA screening.

Effect of glucose oxidase treatment on the aroma qualities and release of cooked off-odor components from heat-treated Hami melon juice

Melon juice produces strong cooked off-odors during heat processing, leading to serious deterioration of aroma quality. In this work, the aroma quality of melon juice, the changes in GOD reaction products, and the interactions of reaction products and cooked off-odor components were analyzed by sensory evaluation, gas chromatography–mass spectrometry/olfactory, ultraperformance liquid chromatography–triple quadrupole mass spectrometry, and isothermal titration calorimetry to study the effect mechanism of glucose oxidase (GOD) on the release of cooked off-odor components from heat-treated melon juice. The results showed that GOD treatment improved the aroma quality mainly by controlling off-odor attributes and maintaining characteristic odor attributes. This was because the reaction products (hydrogen peroxide and gluconic acid) of GOD treatment inhibited the release of cooked off-odor components from heat-treated melon juice through oxidation and hydrophobic effects. Furthermore, these products reduced the loss of characteristic odor compounds by restraining Maillard, degradation, and oxidation reactions during heat processing.

Simultaneous extraction of six phthalic acid esters from polyethylene terephthalate (PET) bottled water using poly (ionic liquid) functionalised silica coated-iron oxide nanoparticles: a risk assessment study

ABSTRACT In this research, a poly (ionic liquid) functionalised silica-coated magnetic nanoparticles were synthesised for the first time and utilised as a novel adsorbent for the simultaneous extraction of six phthalic acid esters from polyethylene terephthalate bottled drinking water samples. The adsorbent was prepared by grafting poly (1-benzyl-3-vinyl-1 H-imidazol-3-ium chloride) onto silica-coated Fe3O4nanoparticles via Cu(0)-mediated reversible-deactivation radical polymerisation method. Various techniques were used to characterise the synthesised nanomaterials. A qualitative analysis was performed using ultra-performance liquid chromatography triple–quadrupole tandem mass spectrometry. Response surface methodology based on central composite design was used to optimise the extraction procedure and the optimised condition for each factor was; pH of3.9, 22 mg of adsorbent, 3.5 min extraction time, 3 min desorption time, 340 µL of elution solvent (methanol).The results of the validation of the method indicated its acceptable accuracy (88.6-100.1%), good linearity (r > 0.995), satisfactory repeatabilities (RSDs ≤ 6% for intra- and inter–day precisions) and high enrichment factors (535–572). The limits of detection and limits of quantification of the proposed method achieved were 1.1-4.6 ngL–1and 3.6-15.3 ngL–1, respectively. In this study, six common PAEs, including dimethyl phthalate, diethyl phthalate, dibutyl phthalate, benzyl butyl phthalate, bis (2-ethylhexyl) phthalate, and di-n–octyl phthalate, were found in PET bottled water within the range of 0.21-0.94 μgL–1, under different storage conditions. Nevertheless, only a negligible risk is caused by the PAEs in PET bottled water for consumers following the recommendations, such as storing at a common place (25°C), in a short period away from the sun.

Tailor-made deep eutectic solvents extraction combined with UPLC-MS/MS determination of icarrin and icarisid II in rat plasma and its comparative pharmacokinetic application

Using green and high efficient solvents to extract and trace active ingredients of traditional Chinese medicine (TCM) in the complex biological samples was still challenging. In this paper, a co-friendly, fast pretreatment method with high extraction efficiency, based on the tailor-made deep eutectic solvent (DES) system, combined with ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-MS/MS) was developed and validated for the determination of icarrin and icarisid II in rat plasma samples, which can be further applied to comparative pharmacokinetic studies after oral administration of Herba Epimedii and icarrin monomer in rats, respectively. PrE (l-proline: ethylene glycol = 1:4 mol/mol) and acetonitrile were optimized and combined as the tailor-made DES at the volumetric ratio of 3:7 to extract icarrin and icarisid II, and to precipitate the protein in rat plasma in one step simultaneously. The extraction efficiency of the tailor-made DES was about 1.7 times of DES (PrE). The extraction recovery of icarrin and icarisid II in rat plasma samples by this method were within the range of 90–110 %, and the lower limits of quantification (LLOQ) were 0.32 ng mL−1 (icarrin) and 0.43 ng mL−1 (icarisid II). There was a linear relationship between 0.32−80.16 ng mL−1 (icarrin) and 0.43−107.4 ng mL−1 (icarisid II), which effectively reduced the detection limit. In this comparative pharmacokinetic study, the maximum plasma concentration (Cmax) and the area under plasma concentration-time curve (AUC0-∞) of two analytes in rat plasma of Herba Epimedii group were both much higher than those in the icarrin monomer group, which suggested that other ingredients in Herba Epimedii may contribute to the in vivo absorption of icarrin and icarisid II. This simple, rapid, relatively green and high effeicient method would provide a new approach for the extraction of active ingredients from complex biological samples.

Development of a Pteris vittata L. compound database by widely targeted metabolomics profiling.

The objective of this work was the development of a detailed, extensive and reliable database of the metabolomes of P. vittata. Using an ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry system (UPLC-QqQ-MS/MS) and based on the knowledge of retention time and mass spectral characteristics of an in-house collection of authentic standards, we screened for the presence of a large collection of natural compounds. The database represents 359 authenticated metabolites, comprising 220 primary and 139 secondary metabolites (70 flavonoids, 16 phenylpropanoic acid derivatives, five coumarins, two stilbenoids, 14 benzoic acids, nine phenols, 20 alkaloids and three terpenoids). Comparison of the accumulation of these compounds in two tissues showed that the aerial parts were enriched in flavonols, whereas the subterranean parts were enriched in anthocyanins. The comprehensive database developed here will be beneficial in improving the understanding of the chemical basis of plant therapeutic profile using multivariate analysis, with a particular example of antioxidant activity.

Screening of a broad-spectrum antagonist-Bacillus siamensis, and its possible mechanisms to control postharvest disease in tropical fruits

Broad-spectrum antagonist Bacillus siamensis (B. siamensis) was screened from ten biocontrol strains because of its excellent antifungal activity on 21 tested pathogens in vitro. In this study, the biological control capability of B. siamensis and its possible action mechanisms on pathogens in postharvest tropical fruits were investigated. Results showed that the volatile organic compounds (VOCs) produced by B. siamensis exhibited significant inhibition to the tested fungi in co-culture. The gas chromatography-mass spectrometry (GC-MS) equipped with solid-phase microextraction (SPME) was adopted for component identification of VOCs, and fifteen diffusible compounds were detected, with properties of inhibiting the growths of hyphae and conidial germinations of fungi proved. Moreover, through Ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QQQ-MS) analysis, the possible existence of surfactin lipopeptide was speculated. The extracellular lytic enzymes, including β-1,3-glucanase, cellulase and protease released by B. siamensis were detected. In vivo, the results of biological control efficacy test showed that B. siamensis suppressed the occurrence of diseases and browning of tropical fruits during storage. Compared with the control, the physiological qualities of postharvest tropical fruits in the treatment were maintained well. The combination of different mechanisms of action led to the broad-spectrum capacity of B. siamensis to defense and attack the pathogens whether in vivo or in vitro. Findings above indicate that broad-spectrum antagonist B. siamensis has great biocontrol potential and could be applied in controlling postharvest diseases in tropical fruits.

Quantification of major royal jelly proteins using ultra performance liquid chromatography tandem triple quadrupole mass spectrometry and application in honey authenticity

Major royal jelly proteins (MRJPs) have been used as endogenous biomarkers for honey authentication. However, no study on quantification of MRJPs in honey authentication was reported. In order to develop a new honey authenticity method based on quantitative analysis of MRJPs in honey, trypsin-digested peptides from MRJPs were examined by mass spectrometry (MS). Three peptides, YNGVPSSLNVISK, TLQMIAGMK, and LTVAGESFTVK were selected as specific markers for MRJP1, MRJP 2 and MRJP 3(MRJP1∼3), respectively. An ultra performance liquid chromatography-triple quadrupole MS (UPLC-TQMS) method used for quantification of MRJPs in honey was developed and validated. A database on the MRJP1∼3 contents of honey established by quantification of 70 authentic samples showed a potential application in honey authenticity identification. The analysis results of 10 commercial honey samples by UPLC-TQMS with reference to the MRJP1∼3 database of authentic honey were compared and confirmed by widely used isotope ratio mass spectrometry, a AOAC Official Method. In conclusion, the results showed our method, a proteomic-based UPLC-TQMS method, has a high accuracy, sensitivity and specificity for honey authenticity.

Rapid and Simultaneously Determination of Six Aristolochic Acids and Two Lignans in Asari Radix Et Rhizoma by Ultra Performance Liquid Chromatography-Triple Quadrupole Tandem Mass Spectrometry

Asari Radix et Rhizoma (AR) is a well-known Chinese herbal medicine which has been used to treat cold headache, toothache, runny nose, rhinorrhea, rheumatism and cough in China. However, the existing Pharmacopoeia standard of AR is still unable to comprehensively evaluating the overall efficacy and safety of AR, whose traditional HPLC detection method has great limitations for the detection of trace aristolochic acids. In order to improving the quality control method of AR, a rapid, simple, and reliable chromatographic analytic method based on ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QQQ-MS) was established for the analysis of 8 target compounds, including 6 aristolochic acids derivates aristolochic acid (AA)-I, AA-II, AA-III, AA-IVa, AA-VIIa, aristololactam I (AL-I), and 2 lignans asarinin and sesamin. Chromatographic separation has been achieved on an ACQUITY UPLC BEH C18 column (50 mm × 2.1 mm, id 1.8 μm) with a mobile phase consisted of water containing 0.1% formic acid (A) and acetonitrile (B) with gradient elution manner. Positive electrospray ionization mode with multiple reaction monitoring (MRM) was applied for the detection of the eight analytes. Method validation results indicated that UPLC-QQQ-MS was precise, accurate and sensitive for quantitative determination of the eight analytes. Finally, the method was successfully used to the analysis of each component in 15 batches of AR herb samples. In addition to existing marker ingredients AA-I and asarinin, considerable amounts of AA-IVa, AA-VIIa, AL-I and sesamin were also found in each AR sample. The result suggesting that AA-IVa, AA-VIIa, AL-I should be limited and sesamin should be added as a quality marker for the quality control of AR.

Removal of pharmaceutically active compounds (PhACs) and bacteria inactivation from urban wastewater effluents by UVA-LED photocatalysis with Gd3+ doped BiVO4

In this study, gadolinium doped bismuth vanadate powders were synthesized, characterized, and tested as a potential photocatalyst for the removal of pharmaceutically active compounds (PhACs) and bacterial inactivation in a real wastewater effluent using UVA as irradiation source. X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) studies revealed that the bismuth vanadate system was successfully doped with 4% of gadolinium in molar mass leading to the formation of a heterostructured photocatalyst.Up to 98.3 % of diclofenac was removed from pure water after 120 min through photocatalysis. However, the photocatalytic performance of the photocatalyst on wastewater effluent was rather variable due to the complexity of the matrix where 22 different PhACs were detected by means of Ultra Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry (UPLC-QqQ-MS/MS). High photocatalytic removal efficiency (80–100%) was observed for some of the studied PhACs (e.g. naproxen and furosemide) whereas moderate efficiency (40–70%) was observed for others (e.g. acetaminophen and azithromycin) after 180 min (UVA dose = 139.5 Wh m−2, t30W = 279 min-1). Some of the studied PhACs like clarithromycin and Ibuprofen showed poor removal efficiency (< 30 %). In disinfection tests, Total coliforms, Escherichia coli, Enterococci, and Klebsiella pneumoniae showed inactivation after direct UVA LED photolysis. Nevertheless, higher inactivation was achieved for Enterococci in the presence of the synthesized photocatalyst showing an increase of 41.1 % in kmax.

Photocatalytic degradation of pharmaceutically active compounds (PhACs) in urban wastewater treatment plants effluents under controlled and natural solar irradiation using immobilized TiO2

The degradation of pharmaceutically active compounds (PhACs) under controlled and natural solar irradiation using immobilized TiO2 was studied in model solution and real urban wastewater effluents. The TiO2 coatings were prepared via the sol–gel method with two different configurations, using simple (thin films) and the “sandwich” approach. Prepared coatings were characterized employing X-ray diffraction (XRD), scanning electron microscopy (SEM), ellipsometry and UV–Vis diffuse reflectance spectroscopy. Decomposition of PhACs detected in urban wastewater effluents (diclofenac, carbamazepine, atenolol, propranolol, albuterol, ofloxacin, ciprofloxacin, azithromycin, erythromycin, hydrochlorothiazide, and furosemide) was monitored using Ultra Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry (UPLC-QqQ-MS/MS). Among detected PhACs three are included in the Watch List (EU Decision 2018/840). The “sandwich” coatings (deposited on frosted support) with the following structure: anatase layer, TiO2 P25 nanoparticles and anatase layer (sealing film) demonstrated the highest photocatalytic activity among all tested TiO2 coatings and were used for elimination of PhACs from real urban wastewater effluents. Photocatalytic treatment of urban wastewater effluents under controlled and natural solar irradiation led to moderate and high removal (>40%) of all detected PhACs except carbamazepine and atenolol, for which poor removal efficiency was observed (~20%). Some of the PhACs compounds present in wastewater effluents, such as diclofenac, ofloxacin, ciprofloxacin, hydrochlorothiazide and furosemide were sensitive to solar photolysis. The efficiency of solar photocatalytic inactivation of Escherichia coli present in urban wastewater effluent was similar to that of solar disinfection (SODIS).

Simultaneous determination of cucurbitacin B, E and I in plasma, urine, and melon and fruit vegetables by ultra performance liquid chromatography-triple quadrupole mass spectrometry using atmospheric pressure chemical ionization

A method for the determination of cucurbitacin B (CuB), cucurbitacin I (CuI) and cucurbitacin E (CuE) in plasma, urine and melon and fruit vegetables was developed by ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). The target analytes in plasma and urine were extracted and cleaned-up by solid supported liquid-liquid extraction, while those in melon and fruit vegetables were extracted with acetonitrile and then diluted with water. CuB, CuI and CuE were separated on an XBridge BEH C18 column (100 mm×3.0 mm, 2.5 μm) with gradient elution using mobile phases of methanol and 0.025% (v/v) ammonia aqueous solution. An atmospheric pressure chemical ionization interface was used as the ion source and the analysis was performed in negative ionization multiple reaction monitoring (MRM) mode. The cucurbitacins in plasma and urine were quantified by the matrix working standard curve internal standard method, while those in melon and fruit vegetables were quantified by the solvent standard curve external standard method. Oleandrin was used as the internal standard. The average recoveries were 89.0%-113% for the three cucurbitacins in plasma and urine, with RSDs of 1.7%-12.2% (n=6). The average recoveries were 87.6%-114% for the three cucurbitacins in melon and fruit vegetables, with RSDs of 4.1%-11.1% (n=6). The limit of detection (S/N=3) of the three cucurbitacins was 0.03 μg/L in plasma and urine, and 5-10 μg/kg in melon and fruit vegetables. The method is simple, sensitive and accurate. It has been used for the determination of cucurbitacins in bitter bottle gourd and in the plasma and urine of patients poisoned by bitter bottle gourd, CuB was successfully detected.